Novo Nordisk A/s

Novo Nordisk Biotech, Inc.

Novo Nordisk-a/s Novo Alle

Novozymes Pharma Biotec A/s

Novozymes A/s

Novozymes, Inc.

Novozymes Adenium Biotech A/s

Ejner Bech Jensen

Henrik Dalbo Slashedge

Karen Gjesing Welinder

Allan Svendsen

Peter Kamp Hansen

Joel R Cherry

Ib Groth Clausen

Ib Jonassen

Poul Erik Pedersen

Per Holse Mygind

Susan L Elrod

Hans-Henrik Kristensen Høgenhaug

Hans-Henrik Kristensen Högenhaug

The invention relates to the use of co-expression of an antimicrobial agent and an enzyme, with a view to improving yield and/or overall production economy. Examples of antimicrobial agents are antimicrobial peptides such as lactoferricins and antimicrobial enzymes such as lysozyme and glucose oxidase, and examples of enzymes are endoglucanase, xylanase, phytase, protease, galactanase, mannanase, dextranase, alpha-galactosidase, pectate lyase, alpha-amylase and glucoamylase. A fusion product comprising the antimicrobial agent and the enzyme and a cleavable linker is novel, and can be used in animal feed and animal feed additives. The invention also describes the use of a protection domain wherein at least 50% of the amino acid residues comprised in the peptide protection domain are D (Asp) and/or E (Glu). The protection or quenching domain serves to temporarily and reversibly inactivate the antimicrobial peptide during the expression.

Recombinant production of antimicrobial peptides

The present invention relates to methods for producing heterologous heme proteins extracellularly comprising transforming a filamentous fungus with a vector comprising a DNA sequence encoding the heterologous heme protein and a DNA sequence encoding a preregion permitting secretion of the expressed heme protein, and culturing the transformed filamentous fungus in a suitable culture medium to produce the heme protein.

Process for producing heme proteins

The present invention relates to methods of producing a polypeptide, comprising: (a) introducing into a respiratory-defective mutant of a cell (i) one or more first nucleic acid sequences which complement the respiratory defect and (ii) a second nucleic acid sequence which encodes the polypeptide; (b) cultivating the cell containing the first and second nucleic acid sequences in a culture medium under aerobic conditions suitable for expression of the first and second nucleic acid sequences; and (c) isolating the polypeptide from the cultivation medium of the cell. The present invention also relates to methods for disrupting a gene in a respiratory-deficient mutant cell. The present invention further relates to respiratory-deficient mutant cells and methods for obtaining such mutant cells.

Methods for producing polypeptides in respiratory-deficient cells

DNA molecule and process for producing insulin precursors having the formula B(1-29) -H.sub.1 --X.sub.2 --Y.sub.2 --Y.sub.1 --A(1-21), wherein B(1-29) are the 29 first amino acid residues of the B chain of human insulin starting from the N-terminus, A(1-21) are the 21 amino acid residues of the A chain of human insulin, X.sub.1 represents a peptide bond or one naturally-occurring alpha-amino acid acid residues, X.sub.2 represents Glu or Asp, and Y.sub.1 and Y.sub.2 each represent Lys or Arg, the positions A6 and A11, A7 and B7 and A20 and B19, respectively, are connected through sulphur bridges, and, if desired, one or more of the glutamic acid residues in positions A4, A17, B13 and B21 are substituted by another naturally-occurring alpha-amino acid residue having an uncharged side chain, are provided. The insulin precursors are prepared by culturing a yeast strain transformed with a replicable plasmid comprising a DNA sequence encoding an insulin precursor of the above formula in a suitable medium and isolating the insulin precursor thus formed from the culture medium. The insulin precursor can be converted into human insulin or insulin analogues by enzymatic treatment in a manner known per se.

DNA sequences encoding insulin precursors and methods of production

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Virum, Denmark

Ejner Bech Jensen